CN102215821A - Methods for preparing polymer microparticles - Google Patents

Abstract

The present invention describes methods and tools for preparing a population of monodisperse polymer microparticles, which are of particular interest in the field of drug delivery.

Description

Be used to prepare the method for polymer micropellet

Technical field

The present invention relates to be used to produce particularly be suitable for single method and apparatus that disperses microparticle population (population) that medicine is sent.

Background technology

In many drug delivery applications, require strictness control to particle size and particle size distribution.An example is from the catheter delivery drug release particles, more specifically for requiring to be delivered to such as the situation in the capillary bed of the tumor of liver tumor.In this drug delivery applications, granule is considered to thromboembolism in minimum capillary tube, and these granules can discharge such as the medicine of cytostatics or partial β or gamma-radiation are provided from minimum capillary tube.For the granule that is loaded with the holmium acetylacetone,2,4-pentanedione, that's how things stand.Particulate size plays an important role in therapeutic efficiency.Too little as fruit granule, they circulate in other zone of for example lung and accumulate.Too big as fruit granule, they will not arrive minimum capillary tube.Therefore utilize granule accurate and evenly customization will realize the most effective therapy.

A kind ofly obtain very from biodegradable polymer that the particulate mode of strict difinition is to use such technology, promptly, liquid with dissolved polymers and medicine is pushed through capillary tube, for example by using the frequency from piezoelectric element or similar device, liquid jet resolves into drop thus.This system is at US6, and 669,916, US6,998,074 and WO2006/003581 and Berkland etc. (2001) J.Control.Release 73, describe among the 59-74.At US6, in 669,916, spray and be accompanied by additional downward power or acceleration so that can produce the drop littler than nozzle diameter.The example of this downward power is the cocurrent flow of poly-vinyl alcohol solution.US6, the problem of mentioning in 669,961 is to produce granule from diameter less than the nozzle of 30 μ m, this nozzle blocks easily.US6, disclosed technology allows to use quite spissated solution (5% polymer) to produce the granule in the desired size scope in 669,961.Yet owing to need additional power, it is complicated more that this production system becomes.Moreover less than the granule of 50 μ m, distribution of sizes is wider than the granule of about 50 μ m for significantly.Similarly current-carrying has been disclosed in US6, and 599,627, this current-carrying is intended to seal and for example contains water (aqueousphase), rather than stream is resolved into drop.This technology produces bigger, promptly greater than the bladder of 100 μ m.

The granule that is of a size of 65 μ m used as Radulescu etc. (2003, Proc.11 ^ThInt.Symp.Control.Rel.Soc) nozzle of drippage as required of described 65 μ m is produced.Herein, aux. pressure can be employed to improve speed of production.From the US6 of same seminar, 998,074 have described the ink ejecting method of drippage as required that uses no aux. pressure, and nozzle is immersed in the liquid to make polymer microballoon.

WO2006/003581 has described the granule littler than granule known in the art and can use and immerse nozzle (this immerses and has used frequency on the nozzle), preferably combines with aux. pressure and produces.The careful contraction of the emulsifying agent drop that is sprayed obtains little granule to 2 μ m.Single disperse, hollow bladder also can as

(Colloids and Surfaces 289 obtains described in 96-104) Deng (2006).In described system, aux. pressure not only allows higher injection rate, and prevents the obstruction of device nozzle.If do not use additonal pressure, will be deposited between fluid to be sprayed and the continuous phase at the interface such as the polymer of polylactic acid.The polymer concentration that the technology described among the WO2006/003581 is used is lower than US6, and the technology of describing in 669,961 or US6,767,637 causes the emulsifying agent drop to be retracted to smaller szie.

The additive of method utilization such as the polyvinyl alcohol of describing in all prior art documents of above quoting comes stabilising emulsifier drop and resulting granules.Add polyvinyl alcohol and two effects are arranged prepare polymer beads from biodegradable polymer.It is stabilising emulsifier drop but also stable by its granule that forms not only.

Except size and uniformity matters, another problem in the ink-jet granule is to receive the fragility of emulsifying agent drop in the fluid.During solvent extraction, stir and very mildly to carry out, otherwise drop is broken easily.

Deng (2006) Colloids ﹠amp; Surfaces, Physicochem.Eng.Aspects 289, and 96-104 has described a kind of system, and wherein microparticle is not stirring after nozzle discharges.

Summary of the invention

Concrete and preferred aspect of the present invention provides in appended independence and dependent claims.From the feature of dependent claims can with the feature of independent claims and with the feature of other dependent claims be only as clearly providing in the claim, to make up rightly and not.

One aspect of the present invention relates to a kind of equipment that is used to produce polymer beads, and described equipment comprises:

-the first reservoir (2) is used to keep emulsifying agent (3),

-the second reservoir (4) is used for keeping receiving fluid (5),

-jet module has at least one nozzle (6), and when this second reservoir (4) when being filled, this at least one nozzle (6) is immersed in this reception fluid (5), allow drop is directly injected in this reception fluid (5),

-inclined surface (7), be positioned in this second reservoir (4) or be associated with this second reservoir (4), make drop when in this reception fluid (5) that is ejected into from this nozzle (6) this second reservoir (4) along this inclined surface (7) motion, wherein this inclined surface (7) have vertically begin and with between 10 degree to gradual change slope that the angle between 45 degree finishes.

On the other hand, the present invention relates to wherein to utilize this equipment, be used to prepare the method for polymer beads.

In a particular aspects, the present invention relates to be used for the method for the single disperse water gelatin polymer microparticle of preparation size between 1 μ m and 100 μ m, wherein said microparticle comprises bioactivator, has wherein utilized according to equipment of the present invention, and described method comprises the steps:

A) preparation comprises the emulsifying agent of the polymer that the hydrogel of described bioactivator forms,

B) use aux. pressure and applying frequency, flow the drop of the emulsifying agent of intravital this nozzle (6) ejection step (a) from being immersed in moisture reception, and,

C) by allowing this drop one time period of motion in this receives fluid, this time period guarantees that described drop removes and desolvates and water comes saturated, with this liquid drip expansion with harden into microparticle, wherein in step (c), this expansion and sclerosis are to carry out by allowing this granule to move downward from the inclined surface on slopes with at least 10 degree in this reception fluid.

In an embodiment of the method for this aspect this according to the present invention, the concentration of polymer is between 1-3%, and the diameter of nozzle is between 1-50 μ m, and the time period that drop moves in receiving fluid is between 2 seconds and 60 seconds.

In the specific embodiment of these methods, step (c) is guaranteed not stir to receive flows intravital microparticle.

In the other specific embodiment of these methods, expansion in the step (c) and sclerosis are to carry out by allowing granule to move downward from the inclined surface on slopes with at least 10 degree in receiving fluid.

In an embodiment again of these methods, inclined surface has the gradual change slope.

Another aspect of the present invention relates to and is used for preparation between 1 μ m and 100 μ m; the method of the monodisperse polymer microparticle between 1 μ m and 50 μ m more specifically; wherein these microparticles comprise bioactivator; this method comprises the steps: the emulsifying agent of the solution of (a) preparation bioactivator and polymer; (b) use aux. pressure and applying frequency; from being immersed in the granule or the drop of the emulsifying agent that receives the intravital nozzle ejection step of stream (a); and (c) by allowing granule in receiving fluid, on inclined surface, to move; these granules or drop are hardened into microparticle; wherein the slope of inclined surface be the gradual change slope and wherein this slope each point have at least 10 the degree, more specifically at least 20 the degree angles.

In the specific embodiment of these methods, polymer is an aquogel polymer.In those embodiment of the method aspect this according to the present invention, wherein employed polymer is an aquogel polymer, can imagine other specific embodiment, wherein step (c) comprise the steps: to allow this granule in receiving fluid on inclined surface one time period of motion, this time period guarantees to remove and desolvates and water comes saturated.

In the specific embodiment of said method, thereby the concentration of aquogel polymer is conditioned and obtains diameter greater than 40% in the emulsifying agent, more specifically greater than the hydrogel particle of 65% nozzle diameter.

In another specific embodiment of said method, the concentration of aquogel polymer is lower than 3% in the emulsifying agent.

In another embodiment of method of the present invention, the concentration of aquogel polymer is between 0.5% to 1.5% in the emulsifying agent, and nozzle has the diameter between 10 μ m and 40 μ m.

In the specific embodiment of all methods of the present invention, bioactivator is hydrophobic.

In other embodiment of method of the present invention, bioactivator is that hydrophilic bioactivator and said method step (a) comprise the steps: to prepare the aqueous solution that contains hydrophilic bioactivator and the water-in-oil emulsifier of polymer solution.

In the specific embodiment of method of the present invention, microparticle has the size between scope 10 μ m and 20 μ m.

In the specific embodiment of method of the present invention, polymer is poly-(ethylene oxide,1,2-epoxyethane) terephthalate and poly-(1,4 butanediol) terephthalate (PEGT/PBT) copolymer.

Use therein in the specific embodiment of the inventive method of inclined surface, the length of inclined surface and slope are arranged such that the time period of moving from the granule of nozzle ejection along inclined surface between 2 seconds to 60 seconds.

In the other specific embodiment of the inventive method, this method is included in step (c) additional step (d) afterwards, wherein this granule further sclerosis by stirring.

In the specific embodiment of the inventive method, nozzle has the diameter less than 30 μ m.

In the specific embodiment of the inventive method, polymer is that aquogel polymer and this method are not exist under the situation of stabilizing agent to carry out in receiving fluid.

Another aspect of the present invention provides size between the monodisperse polymer microparticle population between 1 μ m and 50 μ m between 1 μ m and the 100 μ m and more specifically, and these granules comprise one or more bioactivators.These populations can obtain by said method.

In the specific embodiment of monodisperse polymer microparticle population of the present invention, polymer is an aquogel polymer, and has diameter within the meansigma methods 0.5 μ m of population greater than 90% microparticle.

In the specific embodiment of monodisperse polymer microparticle population, aquogel polymer is the PEGT/PBT copolymer.

In the other specific embodiment of monodisperse polymer microparticle population of the present invention, the granule greater than 90% is within 4% number average diameter.

In the specific embodiment of monodisperse polymer microparticle population provided by the invention, these microparticles comprise hydrophilic bioactivator or comprise hydrophobic bioactivator.

The present invention comprises also that use above describes in detail singly disperses the microparticle population to be used for medicine to send.The present invention also provides and comprises the single drug ingredient that disperses the microparticle population that above describes in detail.

In the specific embodiment of equipment of the present invention, finish with the angle between 20 degree and 30 degree on the gradual change slope.

Above-mentioned and other characteristic of the present invention, feature and advantage will become apparent from the following detailed description of carrying out in conjunction with the accompanying drawings, and accompanying drawing illustrates principle of the present invention by the mode of example.This description only is for the former of example thereby provides, and do not limit the scope of the invention.The reference diagram of hereinafter quoting is meant accompanying drawing.

Description of drawings

Fig. 1 illustrates illustrating of equipment (1) according to an embodiment of the invention, this equipment (1) comprises first reservoir (2), comprises or is connected to second reservoir (4) of inclined surface (7), and this inclined surface (7) is connected to collecting tank (collection bath) (8).Equipment further comprises nozzle (6), and this nozzle arrangement one-tenth makes emulsifier solution drop in from nozzle (6) and drops on when extruding on the inclined surface (7), from the downward slides/rolls in this surface, arrive collecting tank up to them.In the method for the invention, using nozzle (2) will be incorporated into emulsifying agent (3) in first container (2) is expelled to become in second container (4) and receives fluid (5).Alternatively, the vertical wall (9) in second container (4) allow at first with nozzle offset within first compartment/position of top is being provided with frequency, pulse amplitude and pressure, and drop does not arrive collecting tank (8).When parameter is set up, nozzle can be displaced within second compartment with keeping simultaneously immersing/position of top, this second compartment is connected to/comprises inclined surface (7).In the specific embodiment of method of the present invention, the emulsifier solution that sprays from nozzle (6) is dropped on the inclined surface (7), and emulsifier solution arrives the collecting tank (8) of filling with fluid (10) after dropping in and sliding about 2 seconds on this inclined surface.Emulsifying agent drop to lower slider on inclined surface is illustrated.

Fig. 2 illustrates illustrating according to the specific embodiment of equipment of the present invention (1), this equipment comprises first reservoir (2), comprises or is connected to second reservoir (4) of the inclined surface (7) (for example form of tube) with gradual change slope, and this inclined ramp is connected to collecting tank (8).Equipment further comprises nozzle (6).In the specific embodiment of method of the present invention, the emulsifying agent drop that is sprayed from first reservoir (2) that comprises emulsifying agent (3) by nozzle (6) is had the inclined surface guiding on gradual change slope towards collecting tank.The slope of inclined surface (being described as the tube with curvature) vertically begins, and ends at collecting tank (8) with level into about the angle of 20 degree.

Fig. 3 illustrates the particulate distribution of sizes for preparing according to the following specific embodiment that immerses ink-jet: from (poly-(ethylene oxide,1,2-epoxyethane) terephthalate and poly-(1 of 1% PEG-PBT dichloroethanes, 4 butanediols) terephthalate) solution, from 20 μ m nozzles four kinds of different frequency (open circles: 10kHz; Triangle: 12kHz; Filled circles: 17kHz; Square: as 27.6kHz) to immerse ink-jet.

Fig. 4 illustrates the particulate distribution of sizes (solid squares) for preparing according to the specific embodiment that immerses ink-jet from 1% PEG-PBT solution dichloromethane and by immersing the distribution of sizes (open circles) that primary emulsion that ink-jet contains 1%PEG-PBT, 1% water and 98% dichloromethane obtains.

Fig. 5 illustrates according to the granule (left drawing board) of the specific embodiment preparation of immersing ink-jet from 1% PEG-PBT solution dichloromethane and by immersing the SEM photo of the distribution of sizes (right drawing board) that primary emulsion that ink-jet contains 1%PEG-PBT, 1% water and 98% dichloromethane obtains.The bar of bottom, right side is represented 50 μ m.

Fig. 6 illustrate according to from dichloromethane, 1% the PEG-PBT solution that do not add stabilizing agent immerses the particulate SEM photo of the specific embodiment preparation of ink-jet.The bar of bottom, right side is represented 50 μ m.

Fig. 7 is illustrated under 10% and 50% amplification, uses the emulsified water the combine fluorescin fluorescence micrograph from the injection microparticle of 1% PEG-PBT formulations prepared from solutions dichloromethane.

Fig. 8 is illustrated in the dichloroethanes distribution of sizes (grey color triangle) that ink-jet obtains of immersing of immersing particulate distribution of sizes (dark circles) that ink-jet prepares and primary emulsion by containing 0.45%plga and 0.35%pla-peo and 0.5% water by the dichloroethanes that contains 0.45% polylactic-co-glycolic acid copolymer (plga) and 0.35% polylactic acid poly oxirane (PLA-PEO).

Fig. 9 illustrates by what obtain from the ink jet printing of dichloroethane solution deposition and contains iodized oil, suffering-2- base 2,3, the distribution of sizes of the PLGA polymer beads of 5-Triiodobenzoic acid.Last drawing board illustrates distribution of sizes, and end drawing board illustrates the global shape of representative particle.

In different figure, identical reference marks refers to same or similar element.

The specific embodiment

The present invention will be described in conjunction with specific embodiments and with reference to certain illustrative, but the invention is not restricted to this and only be defined by the claims.Any reference marks in claims should not be read as limited field.Described figure only is illustrative and not restrictive.In the drawings, for purpose of explanation, some size of component can be exaggerated and not to scale (NTS) is drawn.In this description and claims, use in the occasion that term " comprises ", do not get rid of other element or step.In the occasion of the indefinite article of for example " " that when quoting singular noun, uses or " ", " being somebody's turn to do " or definite article, comprise a plurality of these nouns, unless regulation clearly in addition.

Moreover the term first, second, third, etc. in description and the claim are used to similar components is distinguished, and not necessarily are used for describing in order or time sequencing.The term that should be understood that such use is that the embodiment of the invention interchangeable and described herein can be operated according to the order in addition of described herein or explanation under appropriate situation.

It only is to understand the present invention in order to assist that following term and definition is provided.These definition should not be interpreted to has the scope of understanding less than those of ordinary skills.

Term used herein " microparticle " refers to the granule of diameter between 1 μ m and 100 μ m.For erose granule, diameter is the longest distance between particulate two outer surfaces.

The material that term " bioactivator " refers to any physiology or pharmacological active substance generally or is suitable for surveying.

Term " hydrophilic " is used as the synonym of " water miscible " in this article and is used to describe the tendentiousness of attraction hydrone and the water solublity of increase in this article.If under 25 ℃ under atmospheric pressure, every premium on currency can dissolve 0.2 gram or more reagent (=0.2mg/ml), then hydrophilic bioactivator is considered to water miscible at this paper.Term used herein " the single dispersion " is meant that when referring to a plurality of microparticle the standard deviation of distribution of sizes is less than 10%.

Term used herein " hydrogel " refers to network natural or the synthetic polymer chain, wherein this network expand into a bit when contacting with water, this point since network in covalently or non-covalently in conjunction with the farmland, the physical chemistry attribute of polymer chain prevents further expansion.But depend on the quantity of hydration group in the polymer, dehydrated hydrogel volumetric expansion when hydration, the coefficient of expansion is 3 to 10, typically coefficient is between about 3 and about 4 (being referred to herein as degree of hydration).

The invention provides novel and improved microparticle that medicine sends and the method that is used to produce them of being suitable for.

One aspect of the present invention provides a kind of a plurality of methods that comprise the monodisperse polymer microparticle of one or more bioactivators that are used to prepare.More specifically, the microparticle average-size is 1 μ m to 100 μ m, more specifically between 2-50 μ m, the most particularly between 15-30 μ m.In embodiment more specifically, the microparticle average-size is 10 μ m to 20 μ m.

The method according to this invention comprises that the polymer micropellet of hydrophilic bioactivator uses ink-jet technology to form.Such as US6, the shortcoming of the known ink ejecting method of describing in 669,961 is the fragility of the emulsifying agent drop of ink-jet.Stirring when solvent extraction must very mildly be carried out, otherwise drop is broken easily.

According to the present invention, ink-jet technology is used, and makes the polymer micropellet sclerosis by allowing microparticle to fall by this in liquid (reception fluid).The reception fluid is an aqueous solution, and it can be diluted and can contains nearly 2%, 10% or 20% the additional compound such as salt, surfactant stabilisers, organic compound, perhaps contains other additive.In this initial bubble and/or between hardening period, receive fluid and typically do not stir for example to avoid because granule collides the mechanical damage that causes mutually.By selecting appropriate container height, can guarantee the emulsifying agent drop because gravity decline specific range hardens to specific degrees thus, they can remove and can stir with further sclerosis from container afterwards.

According to one aspect of the invention, be provided for the method and apparatus of ink-jet microparticle, receive the intravital downward inclined surface of stream and contact with being positioned at from the emulsifying agent drop of nozzle ejection by this, and begin expansion when on this surface, rolling or sliding downwards and/or harden.The most particularly, inclined surface has the gradual change slope.In fact, have been found that, roll downwards or slide with the gradual change slope by allowing emulsifier solution to drop in to receive in the fluid, rather than be subjected to gravity and descend, can obtain emulsifier solution and drop in the inhomogeneity monodisperse particles that receives aging specified time period in the fluid and have increase.

The length of inclined surface can change and will for example determine by the size of drop.Similarly, the structure on slope and the material that is used for this slope can influence and expand and/or hardened particulate performance, and the roughness of its material and hydrophobicity have influence and can be regulated by the technical staff and move downward the required time with further control drop at inclined surface for instance.In specific embodiment, the deceleration gradually that the length on surface and inclination angle (inclination) thereof are designed to guarantee the emulsifying agent drop makes the emulsifying agent drop consuming time between 2 seconds and 60 seconds by moving of rolling or slip along the slope.In specific embodiment, select granule wherein consuming time less than 10 seconds or even less than 5 seconds condition along moving of slope.In specific embodiment, the length of inclined surface is envisioned between 1cm and 2cm, and is perhaps bigger.

In specific embodiment, the slope of inclined surface is fixed.More specifically, the slope on surface is between 10 degree and 45 degree, between 10 degree and 30 degree, perhaps between 20 degree and 40 degree.Inclined surface can be linear, for example the straight tube-like thing.Typically inclined surface extends to from the position (allowing drop to contact or fall on inclined surface with inclined surface) below the nozzle and is used to collect and further alternatively second container of handling the microparticle that is obtained.The opening of nozzle and the distance between the inclined surface (perhaps drop arrives the time that inclined surface needs from nozzle) are included in the best sclerosis/expansion distance (expansion time) of imagining at method of the present invention.Inclined surface can completely or partially place injection nozzle to be immersed in wherein the container, perhaps can be associated with this container in the following manner, at least at the particular location of nozzle, from the emulsifying agent drop of nozzle ejection automatically (for example as the result of gravity or by the direct connection between nozzle and the inclined surface) contact with inclined surface.

In specific embodiment, imagination has the inclined surface on gradual change slope.Imagine the inclination angle of keeping at least 10 degree on the gradual change slope simultaneously particularly, more specifically keep at least 20 ° at each point on slope.Typically, the slope of inclined surface changes at least 10 ° that are positioned at inclined surface end (microparticle is collected in for example collecting tank (8) here) gradually from 90 ° (corresponding to vertically falling from nozzle), more specifically at least 20 °.

In specific embodiment, more specifically when imagination had development length surperficial, inclined surface bending or spiral were to create desired length in small size.

Inclined surface can be smooth or can comprise that curved edge falls this surface to prevent drop.In specific embodiment, the surface is for being connected to the groove or the tube of collecting tank (8), and this collecting tank contains such as receiving fluidic fluid (10).

After rolling downwards or slide from inclined surface, the emulsifying agent drop hardens fully/expands and can stir by rule alternatively and further handle.Therefore, drop can be collected in the collecting tank (8).Additional or alternatively, drop can take out and restores from receiving fluid.

Passive treatment step of the present invention has the advantage of evading institute's liquid droplets damage, and is to implement under the additive situation that does not exist such as the polyvinyl alcohol that typically is used for the stabilising emulsifier drop or the microparticle that forms in specific embodiment.

It is to make us interested especially that Method and kit for of the present invention is used for the microparticle that bioactivator sends in generation, thereby comprises one or more bioactivators alternatively.

According to one embodiment of the invention, tool and method of the present invention is used to produce the microparticle that comprises hydrophobic bioactivator.According to this embodiment, hydrophobic bioactivator uses polymer mixed to become organic solvent with the preparation emulsifying agent together, this emulsifying agent subsequently the method according to this invention by ink-jet.The method of the emulsifying agent of preparation polymer and water-repelling agent is known for technicians.

According to another embodiment of the present invention, Method and kit for of the present invention is used to produce and comprises microparticle hydrophilic or the water-soluble biological activating agent.The polymer micropellet that comprises hydrophilic bioactivator uses dual emulsifying technology to form.At first step, hydrophilic bioactivator is dissolved in the aqueous solution and with the solution of encapsulation polymer in first solvent and mixes to form the primary emulsion of Water-In-Oil (water-in-oil), first solvent and water do not dissolve each other or have limited solubility (less than 5% or 2%) in water, are 1.3% dichloromethane such as dissolubility in water.Primary emulsion adds moisture second solvent to form time emulsifying agent and to continue to homogenize.First and second solvents are selected as making win solvent and second aqueous solvent not dissolve each other or partial miscibility, but the polymer and second solvent do not dissolve each other.After introducing water-in-oil emulsifier, first solvent will be moved in second solvent.Obtain polymer micropellet in this manner, this polymer micropellet contains a plurality of cracks that comprise the water-soluble biological activating agent.Carry out solvent evaporation or extraction subsequently.According to the present invention, tool and method is provided, can obtain the microparticle that uniformity increases by this.In Method and kit for of the present invention, the microparticle that contains aqueous phase is fallen on the inclined surface (6) that is positioned at moisture reception fluid (4) from nozzle (1).More specifically, as mentioned above, inclined ramp has the gradual change slope.

Therefore, method of the present invention is conceived to be used to produce the microparticle that comprises hydrophilic or hydrophobic bioactivator.Nucleic acid, carbohydrate and in general, protein and peptide are water miscible or hydrophilic, and use two as mentioned above emulsifying agent technology usually and be incorporated in the microparticle best.Yet, comprise that the dewatering medicament of some peptide (for example when the Most amino-acids in the peptide is carried hydrophobic side chain) can directly be dissolved in the emulsifying agent that is used for ink-jet in the organic solvent with preparation according to the inventive method with polymer.Therefore, the present invention includes the drug ingredient that contains the obtainable microparticle of the present invention.

Although in view of the vulnerability of protein and peptide, this method and instrument are particularly useful for making the polymer that loads with protein and peptide, also can imagine with the material beyond protein and the peptide certainly and come loadable polymer.For instance, also can be envisioned for the bioactivator of micromolecule, lipid, lipopolysaccharide, polynucleotide and antisense nucleotide (gene therapeutic agents).Thereby such bioactivator that can be combined comprises non-peptide, nonprotein medicine.But the medicine of conjugated polymer character within the scope of the invention also can be in conjunction with having less than 1500 or even less than 500 the medicine than small-molecular weight.

Therefore, in the context of the invention, be conceived to comprise any chemical compound with treatment or preventive effect as the chemical compound of bioactivator.It can be influence or the chemical compound that participates in tissue growth, cell growth, cell differentiation; Can arouse chemical compound such as the biological behaviour of immune response; The chemical compound of perhaps in one or more biological processes, playing the part of any other role.The non-limiting tabulation of example comprises that antimicrobial (comprises antibacterial, antiviral agent and antifungal), antiviral agent, antitumor agent, thrombin inhibitor, anticoagulant, thrombolytic agent, fibrinolytic agent, the vasospasm inhibitor, calcium channel blocker, vasodilation, hypotensive agent, antimicrobial, antibiotic, the inhibitor of surface glycoprotein receptor, anti-platelet agents, antimitotic agent, the microtubule inhibitor, the secretion inhibitor agent, the actin inhibitor, reinvent inhibitor, antimetabolite, antiproliferative (comprising anti-angiogenic agent), anti-cancer chemotherapeutic agents, anti-inflammatory steroids or on-steroidal antiinflammatory, immunosuppressant, the growth hormone receptor antagonist, somatomedin, the dopamine activator, radiotherapy dose, the extracellular matrix composition, ACE inhibitor, free radical scavenger, chelating agen, antioxidant, anti-polymerase and optical dynamic therapy agent.

As above indicated, the particular compound group that can be used for loading according to polymer of the present invention is formed by peptide and protein, in principle can be in conjunction with the peptide and the protein of any kind according to the present invention.Less relatively peptide can be referred to (for example two, three, tetrapeptide) by amino acid whose number.The peptide that has relatively than the amido link (50 aminoacid of as many as) of peanut also can be described as oligopeptide, and the peptide (more than 50 aminoacid) with relatively large number can be described as polypeptide or protein.Except being the polymer of amino acid residue, some protein can further be characterized by so-called quad arrangement, this quad arrangement is the aggregation of many polypeptide, these polypeptide not necessarily come chemical bonds by amido link, but come combination by the power such as electrostatic force and Van der Waals for that the technical staff knows usually.Term peptide used herein, protein or its mixture comprise all above-mentioned probabilities.

Usually, protein and/or peptide are selected based on its biologic activity.Depend on the type of selected polymer, handle the controlled release that obtainable product is very suitable for protein and peptide by this.In specific embodiment, protein or peptide are somatomedin.

Can advantageously be included in peptide or the protein in the loadable polymer or comprise peptide or other example of proteinic entity includes but not limited to immune peptide or immune protein, it includes but not limited to following:

Toxin is such as diphtheria toxin, diphtherotoxin and tetanus toxin.

Viral surface antigen or part virus, such as adenovirus, Ai Baisitan-epstein-Barr virus, hepatitis A virus, hepatitis B virus, herpesvirus, HIV-1, HIV-2, HTLV-III, influenza virus, Japanese encephalitis virus, Measles virus, papillomavirus, paramyxovirus, poliovirus, rabies, virus, rubella virus, cowpox (variola) virus and yellow fever virus.

Bacterium surface antigen or part antibacterial are such as bordetella pertussis, helicobacter pylori, clostridium tetani, diphtheria corynebacterium, escherichia coli, hemophilus influenza, Klebsiella, legionella pneumophilia, Mycobacterium bovis, Mycobacterium leprae, Mycobacterium bovis, gonococcus, Neisseria meningitidis, Bacillus proteus species, bacillus pyocyaneus, Salmonella, dysentery bacterium, staphylococcus aureus, micrococcus scarlatinae, vibrio cholera and plague bacillus.

Cause the parasitic surface antigen or the part parasite of disease, such as Plasmodium vivax (malaria), Plasmodium falciparum (malaria), avette (malaria), plasmodium (malaria), the special sieve pick up (leishmaniasis) of leishmania, leishmania (leishmaniasis), leishmania brasiliensis (leishmaniasis), Trypanosoma rhodesiense (sleeping sickness), Bu Shi gambian trypanosomiasis (sleeping sickness), Ku Shi trypanosomicide (South American trypanosomiasis), Schistosoma mansoni (schistosomicide), Schistosoma haematobium (schistosomicide), Schistosoma japonicum (schistosomicide), trichinella (trichonematosis), duodenum strongyloides intestinalis (ancylostome), duodenum ancylostoma caninum (ancylostome), Necator americanus (ancylostome), Filaria philippinensis (filaricide), Wuchereria malayi (filaricide), Loa loa (filaricide), Acanthocheilonema perstans (filaricide), guinea worm (filaricide), and filaria volvulus (filaricide).

Immunoglobulin is such as IgG, IgA, IgM, anti-rabies immune globulin and anti-vaccinia immune globulin.

Antitoxin is such as botulinum Antitoxin, diphtheria antitoxin, Gas Gangrene Antitoxin, tetanus antitoxin.

Cause antigen at the immune response of hand-foot-mouth disease.

Hormone and somatomedin are such as follicule-stimulating hormone (FSH), prolactin antagonist, angiogenin, epidermal growth factor, calcitonin, erythropoietin, throtropin releasing hormone, insulin, HGH, type-1 insulin like growth factor and 2, the skeletal growth factor, human chorionic gonadotropin, metakentrin, nerve growth factor, thyroliberin (ACTH), luteinising hormone-releasing hormo (LHRH), parathyroid hormone (PTH), throtropin releasing hormone (TRH), vassopressin, cholecystokinin, and corticotropin releasing hormone; Cytokine, such as interferon, interleukin, colony stimulating factor, tumor necrosis factor: fibrinoclase, such as urokinase type plasminogen activator kidney; And thrombin, such as protein C, Factor IX, factors IX, factor VII and Antithrombin III.

The example of other protein or peptide is an albumin, atrial natriuretic peptide, feritin, superoxide dismutase, alpha1 Anti-trypsin, Curosurf protein, Bacitracin, bestatin, ciclosporin, sleep inducing peptide (DSIP), endorphins, glucagon, Gramicidin, melanocyte inhibitory factor, neurotensin, oxytocin, somatostatin, teprotide, serum thymic factor, thymosin, Desmopressin, dermorphin, met-enkephalin, Peptidoglycan, full element, Thymopentin, fibrin degradation product (FDP), des enkephalin-alpha-endorphin, gonadotropin releasing hormone, leuprorelin, α-melanocyte-stimulating hormone and metkefamide.

Antitumor agent replaces group, D actinomycin D, thioguanine, daunorubicin, treosulfan, amycin, D2EHDTPA amide, estramustine, vinblastine, etoglucid, vincristine, etoposide, vindesine and paclitaxel such as altretamine, fluorouracil, amsacrine, hydroxyurea, asparaginase, ifosfamide, bleomycin, lomustine, busulfan, melphalan, chlorambucil, Mercapto base purine, chlormethine, methotrexate, cisplatin, mitomycin, cyclophosphamide, procarbazine, cytosine arabinoside, teniposide, dacarbazine, plug.

Antibacterial comprises:

Antibiotic is such as ampicillin, NAFCILLIN, the amoxicillin, oxazacillin, the azlocillin, benzylpenicillin, Carbenicillin, penicillin V, dicloxacillin, the phenethicillin, the flucloxacillin, piperacillin, mecillinam, the sulbenicillin, the methicillin, ticarcillin, the mezlocillin, cephalosporin: cefaclor, cephalosporin, cefadroxil, cefapirin, cefamandole, cefradine, rocephin, cefsulodin, cefazolin sodium, ceftazidime, ceforanide, rocephin, cefoxitin, cefuroxime, cefacetrile, latamoxef, and cefalexin.Aminoglycoside such as amikacin, neomycin, dibekacin, kanamycin, gentamycin, netilmicin, kanamycin, tobramycin.Macrolide such as amphotericin B, novobiocin, bacitracin, nystatin, clindamycin, polymyxin, colistin, spiramycin, erythromycin, spectinomycin, lincomycin, vancomycin.Tetracycline such as chlortetracycline, oxytetracycline, demeclocycline, Rolitetracycline, doxycycline, tetracycline, minocycline.Other antibiotic such as chloromycetin, rifamycin, rifampicin and thiamphenicol.

Chemotherapeutics is such as sulfa drugs sulfadiazine, sulfamethizole, sulfamethazine, bactrim, sulfadimidine, sulfamethoxypyridazine, bacteresulf, sulfaphenazole, sulfalene, sulfasomidine, sulfamethyldiazine, bacteresulf with have bactrim or the trimethoprim of sulfametrole.

The urinary tract antiseptic is such as methylamine, quinolones (norfloxacin, cinoxacin), nalidixan, nitro compound (nitrofurantoin, nifurtoinol) and Ou Suolin acid.

The medicine that is used for anaerobic infection is such as metronidazole.

Be used for phthisical medicine, such as aminosallcylic acid, isoniazid, cycloserine, rifampicin, ethambutol, thiocarlide, ethionamide and viomycin.

The medicine that is used for leprosy is such as thioacetazone, rifampicin, clofazimine, sulfoxone sodium and diaminourea hexichol (DDS, dapsone).

Antifungal is such as amphotericin B, ketoconazole, clotrimazole, miconazole, econazole, natamycin, flucytosine, nystatin and griseofulvin.

Antiviral agent is such as acyclovir, idoxuridine, amantadine, methisazone, cytosine arabinoside, vidarabine and ganciclovir.

The chemotherapy of amoebiasis is such as chloroquine, diiodohydroxyquinoline (Iodoquinol), clioquinol, metronidazole, dehydroemetine, paromomycin, diloxanide, bran tinidazole and ipecine.

Antimalaric is such as chloroquine, pyrimethamine, oxychloroquine, quinine, mefloquine, sulfanilamide/pyrimethamine, pentamidine, naganol, primaquine, trimethoprim and proguanil.

The anthelmintic agent is such as antimony potassium tartrate, niridazole, sodium antimonate dimercapto fourth, oxamniquine, bepheninum, piperazine, dichlorophen, praziquantel, diethylcarbamazine, Pyrantel Pamoate, hycanthone, povan, levamisole, Stibophen, mebendazole, tetramisole, metrifonate, probenazole and niclosamide.

Antiinflammatory is such as aspirin, mefenamic acid, diclofenac, naproxen, azo acetone, niflumic acid, benzydamine, oxyphenbutazone, diclofenac, piroxicam, fenoprofen, pirprofen, flurbiprofen, sodium salicylate, ibuprofen sulindac, indometacin, tiaprofenic acid, ketoprofen and tolmetin.

The gout agent is such as colchicine and allopurinol.

Act on (Opium) analgesics of nervus centralis, such as alfentanil, methadone, bezitramide, morphine, buprenorphine, nicomorphine, butorphanol, pentazocine, codeine, dolantin, dextromoramide, Piritramide, dextropropoxyphene, sufentanil and fentanyl.

Local anesthetic such as articaine, mepivacaine, bupivacaine, procaine, etidocaine, procaine, lignocaine and tetracaine.

Be used for parkinsonian medicine, such as amantadine, diphenhydramine, apomorphine, profenamine, methylsulfonyl benzatropine, Lergotrile, biperiden, levodopa, bromocriptine, lisuride, carbidopa, methixene, chlorphenoxamine, orphenadrine, cycrimine, procyclidine, dexetimide and benzhexol.

The active muscle relaxant of nervus centralis is such as baclofen, carisoprodol, chlormezanone, chlorzoxazone, cyclobenzaprine, dantrolene, diazepam, febarbamate, mephenoxalone, mephenesin, metaxalone, methocarbamol and tolperisone.

Corticosteroid comprises:

Mineralocorticoid is such as hydrocortisone, desoxycortone and fludrocortisone.

Glucocorticoid is such as beclomethasone, betamethasone, cortisone, dexamethasone, fluocinolone acetonide, fluocinonide, fluocortolone, fluorometholone, fluprednisolone, flurandrenolide, halcinonidedcorten, hydrocortisone, medrysone, methylprednisolone, paramethasone, prednisolone, prednisone and triamcinolone acetonide (An Naide).

Androgen comprises:

The androgenic steroids that is used for the treatment of is such as danazol, fluoxymesterone, mesterolone, methyltestosterone, testosterone and salt thereof.

The desogestrel that is used for the treatment of is such as calusterone, nortestosterone and salt thereof, drostanolone, anavar, ethylestrenol, oxymetholone, methandriol, stanozolol metandienone and testolactone.

Androgen antagonist is such as cyproterone acetate.

The estrogen that comprises the estrogens sterin that is used for the treatment of is such as diethylstilbestrol, estradiol, estriol, ethinylestradiol, mestranol and quinestrol.

Estrogen antagonist is such as chlorotrianisene, clomifene, ethamoxytriphetol, nafoxidine and tamoxifen.

Progestogens is such as allylestrenol, desogestrel, dimethisterone, lynestrenol, lynestrenol, ethisterone, ethynodiol diacetate, ethynodiol diacetate, hydroxyprogesterone, levonorgestrel, lynestrenol, medroxyprogesterone, megestrol, norethindrone, norethindrone, Norethynodrel, methylnorethindron and progesterone.

The thyroid medicine comprises:

The thyroid medicine that is used for the treatment of is such as thyroxine and liothyronine.

The antithyroid drug that is used for the treatment of is such as Athyromazole, thiamazole, methylthiouracil and propylthiouracil.

According to specific embodiment, bioactivator is the label material such as contrast agent or label.Preferably, contrast agent or label are hydrophobic, perhaps have special properties.The preparation of these types can be combined very effectively and be degraded to big degree up to it just can be from polymeric matrix from discharging.Particularly appropriate formulation is the iodate x-ray contrast agent of liquid form.For this purpose, can use product E thiodol for instance, it is iodinating poppy seed oil.For higher iodine load, can synthesis example such as content of iodine greater than suffering-2- base 2,3 of 1000mg/ml, the iodized oil of 5-Triiodobenzoic acid.

Polymer micropellet can comprise one or more bioactivators or can comprise the combination of therapeutic agent and contrast agent.Except water miscible bioactivator, also can be in conjunction with other water soluble compound such as antioxidant, ion, chelating agen, dyestuff, imaging compounds.In addition, except hydrophobic bioactive compound, other chemical compound can be added to the solvent that is used to dissolve this polymer.These hydrophobic compounds can be antioxidant, ion, chelating agen, dyestuff or imaging compounds.As mentioned above, Method and kit for of the present invention can be used to produce and contains hydrophobic and polymer micropellet hydrophilizing agent.Water-repelling agent directly is attached in the polymer solution, and hydrophilizing agent is combined via two emulsifying agent technology.

For the method for this aspect, can use the biocompatible polymeric of any kind according to the present invention.In certain embodiments, biocompatible polymeric also is a biodegradable.The example of biodegradable biocompatible polymeric is polylactic acid, poly-Acetic acid, hydroxy-, bimol. cyclic ester, polylactic acid Acetic acid, hydroxy-, bimol. cyclic ester, polycaprolactone, Merlon, polyesteramide, poly-anhydride, polyamino acid, poe, polydioxanone, polyalkylene alkylates, polyacetals, polybutylcyanoacrylate, Biodegradable polyurethane and solvate and copolymer.The polymer that typically comprises copolymer, its solvate or its mixture of polylactic acid, lactic acid and Acetic acid, hydroxy-, bimol. cyclic ester is used.These polymer can be formed by the monomer of single isomerism type or isomer mixture.

The example of the compatible polymer of abiotic degradable biological is the polymer of polypropylene, ethylene vinyl acetate and other acyl substituted cellulose acetate, non-degradable polyurethane, polystyrene, polrvinyl chloride, polyvinyl fluoride, polyvinyl imidazol, chlorosulfonic acid polyolefin, poly(ethylene oxide) (PEO), its solvate and copolymer.

In specific embodiment, polymer is PLA-PEO (polylactic acid poly oxirane) copolymer.

Another aspect of the present invention relates to the Method and kit for that is used to use ink-jet to prepare and comprises the monodisperse polymer microparticle of bioactivator, this microparticle size is between 1 μ m and 100 μ m, more specifically between 2 μ m and 50 μ m, wherein polymer is a hydrogel.Have been found that and use aquogel polymer to allow to produce granule that this is a problem for previous disclosed technology (for example US6,669,961) less than 30 μ m from low concentration polymer.In liquid, expand and/or during the polymer beads of sclerosis ink-jet, organic solvent diffuses out from granule, have the back-diffusion of water in the microparticle simultaneously.In hydrogel particle formed, the favourable outcome of this back-diffusion was not realized as yet.After the gel aqueous fluid instillation was mapped in the moisture reception fluid, a large amount of water were absorbed by hydrogel particle, cause significant expansion.This expansion does not rely on the time that drop contacts with moisture reception fluid.Equilibrium valve almost obtains at once.When leaving nozzle, drop has the diameter that is slightly larger than nozzle diameter, depends on the amount of applied pressure and institute's spraying polymer.In the practice, produce the drop that diameter is about 45 μ m for for example 30 microns nozzle ejection from diameter.The final diameter of hydrogel particle will depend on the combination of polymer concentration and hydrogel degree of hydration, but wherein the hydrogel degree of hydration is to be determined by the group number of hydration on the polymer.By water the compensate for dilatation of dehydrated hydrogel granule on volume is generally 3 to 4 the factor, but can reaches 5,6,8 or 10 value.

Generally speaking, owing to remove the result desolvate, diameter is that 50 μ m, polymer concentration are the hydrogel drop of 1% to 3% injection, at first with 100 to 33 factor retraction volume, and is shrunk to the granule that diameter is about 11 μ m to 16 μ m on diameter.When these granules increase volume in hydration and with the factor 3, expand into the granule that diameter is about 15 μ m to 22 μ m.When granule increases volume in hydration and with the factor 4, expand into the granule that diameter is about 17 μ m and 25 μ m respectively.Come the type (being degree of hydration) and the polymer concentration of selective polymer according to the desired size of result's hydrated hydrogel polymer beads, this is within technical staff's limit of power.

Aquogel polymer thereby have such advantage can spray the polymer drop with relative smaller size smaller, obtains relatively large microparticle simultaneously after expanding.By using littler drop, the solvent that is present in the hydrogel significantly increases to external diffusion from drop.The expansion of polymer beads as a result and/or the required total time of hardening significantly descend.Moreover owing to receive fluid less by solvent contamination, the more heavy gradient of the solvent between granule and the reception fluid is maintained, and promotes the expansion and/or the sclerosis of polymer beads once more.

Therefore, in specific embodiments of the invention, imagination is used the emulsifying agent with low concentration of water gelatin polymer, and for example 0.5% to 3%, more specifically 0.5% to 2%, such as the aquogel polymer of 1.5% in the emulsifying agent.In other specific embodiment, this emulsifying agent is from nozzle ejection, and nozzle diameter is between 1 μ m and 100 μ m, more specifically between 10 μ m and 50 μ m, even more specifically between 20 μ m and 30 μ m.By regulating nozzle diameter, the controlled expansion of the ink-jet drop of hydrogel emulsifying agent allows to produce such microparticle, its diameter between nozzle diameter 40% and 120% between, more specifically diameter be the nozzle diameter that uses greater than ink-jet 70% to 85% (particularly greater than 75%, greater than 80%, perhaps greater than 85%).In specific embodiment, the diameter of the hydrogel particle that obtains after expanding can be the same with nozzle diameter big or bigger.

Example has partly been demonstrated, and is that about 1% aquogel polymer concentration of the nozzle ejection of 20 μ m can be used for obtaining the polymer beads that diameter is about 17 μ m from diameter, and this diameter is corresponding to 85% nozzle diameter.

As mentioned above, the polymer that uses hydrogel to form is made microparticle, and this allows to use polymer concentration to be lower than 3%, and perhaps even be lower than 2% emulsifying agent, this concentration significantly is lower than the polymer concentration that uses traditionally.This emulsifying agent has lower viscosity, and this is beneficial to ink-jet, because force emulsifying agent to pass through the required pressure of ink discharge device still less.The polymer that uses hydrogel to form also has such advantage, and during ink-jet, nozzle pollutes and reduces.

The present invention thereby be provided for producing and comprise method bioactivator, the aquogel polymer microparticle of size range between 1 μ m and 50 μ m.Typically, method of the present invention comprises step: prepare the emulsifying agent of the polymer of aforesaid hydrogel formation, this emulsifying agent comprises bioactivator; And use aux. pressure and applying frequency, flow intravital nozzle ejection emulsifying agent liquid from being immersed in moisture reception.By allowing drop one time period of motion in receiving fluid, this time period guarantees that described drop removes and desolvates and water comes saturatedly that drop is hardened into microparticle.Typically, greater than 30%, particularly greater than 40%, more specifically greater than 50%, the most particularly between 60% and 98% or more between solvent be removed and all or part ofly substituted by water.In the specific embodiment of the inventive method, granule moves in receiving fluid, does not stir.The most particularly, on inclined surface, slide, granule is moved in receiving fluid by allowing granule as described herein.

Therefore, the invention provides the particulate single population that disperses of aquogel polymer, particle diameter is between 1 μ m and 100 μ m, and more specifically diameter is in the scope between 10 μ m and 30 μ m, the most particularly between 10 μ m and 20 μ m.The granule of size between 10 μ m and 20 μ m particularly is suitable for such as the medical application that forms thromboembolism in the thinnest blood vessel.

According to specific embodiment, the aquogel polymer microparticle comprises one or more bioactivators.In specific embodiment, hydrogel particle comprises hydrophilic bioactivator.Alternatively, bioactivator is that hydrophobic and two emulsifying agent technology are used as mentioned above.The character of bioactivator is not crucial, and the suitable example of contemplated bioactivator is above open.

According to another specific embodiment, the hydrogel microparticle is produced in solvent by ink-jet, and wherein microparticle drops on the inclined surface with gradual change slope, as previously mentioned.

Hydrogel is particularly useful for the controlled delivery of water wetted material particularly, because the water-soluble zone of polymer makes that water can be near the material that be embedded in the polymer.Depend on the characteristic aperture in the polymer, can be by discharging from polymer diffusion from polymer diffusion and/or by material when degrading at material before the degraded, this characteristic aperture is to be controlled by molecular weight between crosslinked and crosslink density.Because the fixing and protectiveness effect of gelinite, the deactivation of embedded material is lowered, and the calamitous effect of the burst that is associated with other controlled release system is avoided.Hydrolysis gradually is beneficial to the final free macromole of controlled release in the body by the terminal ester link in the degeneration of polymer.Generally speaking, hydrogel is formed by the polymerisable macromonomer of biodegradable, and this macromonomer comprises core, the end cap in the extension on each end of core and each extension.Core is hydrophilic polymer or oligomer; Each extends to the biodegradable oligomer; And each end cap is for being cross-linked into oligomer, dimer or the monomer of macromonomer.In another specific embodiment, core comprises molecular weight between about 400Da and 30, the hydrophilic Polyethylene Glycol oligomer between the 000Da; Each extension comprises molecular weight biodegradable poly-('alpha '-hydroxy acids) oligomer between about 200Da and 1200Da; And each end cap comprises acrylic ester monomer or the oligomer (that is, contain carbon-to-carbon double bond) of molecular weight between about 50Da and 200Da, and it can crosslinked and polymerization between copolymer.Another embodiment combines: be about 10 by molecular weight, the core that the Polyethylene Glycol oligomer of 000Da is formed; Be about the extension that the polyglycolic acid oligomer of 250Da is formed by molecular weight; And be about the end cap that the acrylate matrix of 100Da is formed by molecular weight.

The suitable example that is used as the material in the water-soluble zone of core is a Polyethylene Glycol, poly(ethylene oxide), polyvinyl alcohol, polyvinylpyrrolidone, poly-ethyl azoles quinoline, the block copolymer of poly(ethylene oxide) copolymerization expoxy propane is such as the polysaccharide or the carbohydrate of hyaluronic acid, dextran, heparinase, chondroitin sulfate, heparin or alginate, perhaps such as the protein of gelatin, collagen, albumin or ovalbumin.

The biodegradable link that is subject to such as ester, peptide, anhydride, ortho esters and phosphoric acid ester bond can be used in the biodegradable zone, is constructed by polymer or monomer.

The example of hydrolyzable biodegradable composition is the polymer and the oligomer of glycolic, lactic acid, 6-caprolactone, other alpha-hydroxy acid, and produces avirulence or be present in intravital other biodegradable polymer as the homergy product.Suitable poly-('alpha '-hydroxy acids) is polyglycolic acid, poly-(DL-lactic acid) and poly-(L-lactic acid).Other useful material comprises polyamino acid, polyanhydride, poe and poly phosphate.Such as poly-(6-caprolactone), it is poly-that (ε (3 caprolactone), poly-(δ-Wu Neizhi) and the polylactone that gathers (gamma-butyrolacton) they also are useful.

The example is the few lactyl acrylate of PEG.Herein, Polyethylene Glycol or PEG central construct unit (core) are based on its high hydrophilic and water solublity and incidental outstanding bio-compatibility but useful.Weak point poly-('alpha '-hydroxy acids) such as polyglycolic acid is suitable chain extension, because it is hydrolyzed into glycolic and degraded fast by the ester link, glycolic is a kind of harmless metabolite.These networks can be used for embedding and disperse water soluble drug and enzyme and they are sent by controllable rate equably.Other suitable chain extension is polylactic acid, polycaprolactone, poe and polyanhydride.Also can use polypeptide.

In specific embodiment, the hydrogel microparticle is that the copolymer from poly alkylene glycol and aromatic polyester prepares.This copolymer comprises: based on the weight of copolymer, from about 30wt.% to about 99wt.%, first composition of 30wt.% to 90wt.% particularly, this first composition comprises poly alkylene glycol and has the unit of molecular formula for-OLO-CO-Q-CO-, wherein L is the bivalence organic free radical in reservation after polyoxyalkylene ethylene glycol is removed terminal hydroxyl, and Q is the bivalence organic free radical; And based on the weight of copolymer, from about 1wt.% to about 70wt.%, second composition of 10wt.% to 70wt.% particularly, this second composition is aromatic polyester and has the unit of molecular formula for-O-E-O-CO-R-CO-, wherein E is replacement or unsubstituted alkylene or the oxyalkylene group free radical with 2 to 8 carbon atoms, and R is for replacing or unsubstituted bivalence aromatic series free radical.

In one embodiment, poly alkylene glycol is to be selected from Polyethylene Glycol, polypropylene glycol and Polyethylene Glycol and such as its copolymer of poloxalkol.In one embodiment, poly alkylene glycol is a Polyethylene Glycol.Term alkylene and polyalkylene are often referred to acts as for what isomer structure, be that propylene comprises 1,2-propylene and 1, the 3-propylene, butylene comprises 1,2-butylene, 1,3-butylene, 2,3-butylene, 1,2-isobutene., 1,3-isobutene. and 1,4-butylene (Tetramethylene .) and be similar for higher alkylene congener.If desired, the poly alkylene glycol composition with dicarboxylic acids residual-CO-Q-CO-stops so that the coupling with the polyester composition to be provided.Group Q can be the aromatic group that has with the identical definition of R, perhaps can be the aliphatic group such as ethylene, propylene, butylene etc.

In the polyester composition, bivalence aromatic group R can be phenylene, pyridylidene, naphthylene, xenyl, oxygen diphenyl, phenyl disulfide base, methyldiphenyl base, alternatively by four of as many as, two of as many as are selected from following substituent group and replace particularly: chloro, nitro and C1-C4 alkoxyl, and the alkyl of other fluorine-based, hydroxyl and C1-C4.Typically, aromatic series free radical R is not substituted, and R is 1 particularly, the 4-phenylene.The example of polyester-O-E-O-CO-R-CO-comprises polyethylene terephthalate, polypropylene ethylene glycol terephthalate, polybutylene terephthalate (PBT), poly-M-phthalic acid butanediol ester, poly-(ethylene 4,4 '-the diphenyl methylene dicarboxylic ester), poly-( butylene 5,6,7,8-naphthane-1, the 4-dicarboxylic ester) etc.

In specific embodiment, polyester is selected from polyethylene terephthalate, polypropylene ethylene glycol terephthalate and polybutylene terephthalate (PBT).More specifically, polyester is a polybutylene terephthalate (PBT).In unusual specific embodiment, copolymer is Polyethylene Glycol/polybutylene terephthalate (PBT) block copolymer.

Poly alkylene glycol can have from about 200 to about 20,000 molecular weight.The definite molecular weight of poly-(oxygen ethylene) ethylene glycol depends on the various factors that comprises by the bioactivator type of matrix encapsulation.

In specific embodiment, aquogel polymer is the copolymer (PEGT/PBT) of poly terephthalic acid oxirane and poly-(1,4 butanediol) p-phthalic acid.Wherein the PEGT/PBT part by weight can be in from 80/20 to 70/30 to 60/40 scope.In specific embodiment, the Polyethylene Glycol composition of 70/30 and 60/40 copolymer has 1,000 molecular weight.In other specific embodiment, the molecular weight of the Polyethylene Glycol composition of 80/20 copolymer is respectively 1,000,2,000 and 4,000.

Wherein the attendant advantages that has of the inventive method of being used in ink-jet of the polymer that forms of hydrogel is not need the stable additive of using.Generally in the microparticle of preparation biodegradable polymer, need additive.Employed common additives is polyvinyl alcohol (PVA), and it can be with different molecular weight and degree of hydrolysis and commercial acquisition.On aspect the solution of mixed biologic degradable polymer during having the organic solvent of water prepares microparticle, PVA has three kinds of functions.At first, PVA is the colouring stabilizer that goes out that is used for after ink-jet the emulsifying agent drop that forms.Secondly PVA is as the stabilizing agent of hardened microparticle, and its three PVA has influence for the shearing force between the emulsifying agent drop.

The polymer that uses hydrogel to form in specific embodiments of the invention, this has removed the needs of PVA to the Stabilization of emulsifying agent drop stability and microparticle stability.Similarly, no longer need viscosity-increasing agent to control the microparticle size.Spray and controlled expansion by immersing in aqueous solution according to the present invention, do not utilize shearing force to decompose the emulsifying agent drop, so this processing can be carried out under the situation of continuous phase not adding PVA.Therefore need cleaning step still less and handle faster subsequently to cause encapsulation more efficiently.

Method of the present invention allows to use two emulsifying agent technology in addition, wherein when aquogel polymer is used, can omit from the first emulsifying agent step such as the surfactant of the addition polymers (Pluronic) of polypropylene glycol and oxirane.

In ink ejecting method of the present invention and instrument, use nozzle to utilize aux. pressure and piezoelectric device combination such as ink gun, the bioactivator in polymer emulsifier is introduced into fluid (be also referred to as and receive fluid).According to the known ink ejecting method that immerses, nozzle is immersed in and receives in the fluid.

The equipment that immerses ink-jet that is used for microparticle is described at for example WO2006003581 and US6599627.

This equipment typically comprises:

-the first reservoir (2) is used to keep the emulsifying agent (3) of bioactivator and polymer,

-the second reservoir (4) is used for keeping receiving fluid (5), and

-jet module has to be used for emulsifying agent is ejected into and receives stream intravital at least one nozzle (6).

In equipment of the present invention, be used to receive fluidic second reservoir (4) and nozzle (6) and be changed to feasiblely, after filling up second reservoir (4) with the reception fluid, nozzle (6) is immersed in and receives in the fluid.

The diameter of nozzle can depend on regulates the particulate size and/or the employed polymer concentration that obtain, typically between 1-100 μ m.In specific embodiment, equipment of the present invention comprises the nozzle of diameter between 20 μ m and 50 μ m.

In one embodiment, equipment comprises that control system is in 100kHz so that injection is controlled at ^-1To 0.1kHz ^-1Injection rate in the scope.In another embodiment, control system is arranged to spray with the pulse mode operation, and control system is arranged to use square wave form driving pulse to jet module particularly.In another embodiment, spraying system comprises that plurality of nozzles and control system are arranged to regulate drop size at single nozzles.In another embodiment, reservoir is provided with temperature control system.

The specific embodiment of using in the methods of the invention that immerses ink-jet exists

Deng description in (2006) (above quoting).It is that the microdroplet inkjet nozzle (MK-140H) of the Piezoelectric Driving of 50 μ m, 30 μ m and 20 μ m carries out that the experiment of this paper is to use diameter.External pulse generator (Fluke PM5139) is used to obtain the frequency of 20kHz scope.Optionally microdroplet driver (MD-E-201H) also allows acquisition pulse to trigger image to follow a forming process.Nozzle places in the liquid of reservoir.Constant pressure is employed to prevent plug nozzle and to allow big drop to form speed, because the static fluid pressure of black reservoir is insufficient.Typical pressure is at the 0.3bar of 50 μ m nozzles, at the 0.8bar of 30 μ m nozzles and at the 1.6bar of 20 μ m nozzles.(illustrate among Fig. 1) that in one embodiment reservoir contains two compartments of being separated by vertical wall (' 9 '), one of them compartment is used to be provided with frequency and pressure, thereby obtains the emulsifying agent drop array of strict difinition.When reaching this point, nozzle places in second compartment, keeps nozzle ejection simultaneously and is immersed, and this second compartment is the compartment that is used to produce emulsifying agent.After 15 minutes, nozzle is retracted, and pulse generator cuts out.Nozzle removes from solution, and pressure still is employed.Only after nozzle was fully in air, aux. pressure cut off.So doing is not to be back in the nozzle in order to ensure aqueous solution, and this can cause stopping up.Utilization is such as above-mentioned ink discharge device, and drop is injected with the speed of about 5000 to 100.000 drop/seconds.Typically throughput rate is about 25.000 per seconds.

According to one aspect of the invention, such equipment is provided, it allows after ink-jet the specific post processing to microparticle.The equipment of this aspect comprises inclined surface (7) according to the present invention, and it is placed in first reservoir (4) or is related with it, is used for keeping receiving fluid (5).Inclined surface (7) is placed to and makes from the drop contact of nozzle (6) ejection or drop on the starting point on slope of inclined surface (7).As mentioned above, formed microparticle is very fragile and can be owing to for example damage by the active post processing of stirring microparticle in receiving fluid.According to the present invention, such equipment is provided, wherein initiatively post processing is postponed or is omitted.More specifically, equipment is designed to guarantee passive post-processing step, promptly goes up slip at inclined surface (7) by the drop that allows to be sprayed in receiving fluid.More specifically, equipment of the present invention comprises inclined surface, and wherein this inclined surface has between 10 degree and 45 degree, more specifically the slope between 20 degree and 45 degree.According to another embodiment, equipment of the present invention has the gradual change slope, and it vertically begins and finishes to spend to the angle between 45 degree between 10.Example according to the equipment of the specific embodiment of the invention is provided in Fig. 1 and 2.

It is single dispersion that method and apparatus of the present invention allows to produce, and promptly has the very microparticle population of narrow size distribution.Thereby another aspect of the present invention relates to single microparticle population that disperses, and its diameter is 1 μ m to 100 μ m, 2 μ m to 50 μ m more specifically, 10 μ m to 30 μ m particularly, even 10 μ m to 20 μ m more specifically.In this population, the microparticle greater than 90% has the diameter in the meansigma methods 0.7 μ M that drops on described population.In specific embodiment, when aquogel polymer was used, population was to be characterized by such fact the most particularly, and the microparticle greater than 90% has the diameter in the meansigma methods 0.5 μ M that drops on this population.In specific embodiment, in this population greater than 90% microparticle have the average diameter of dropping on 4% in diameter.According to specific embodiment, microparticle is the aquogel polymer microparticle.According to another embodiment, microparticle contains bioactivator.According to another embodiment, microparticle contains hydrophilic bioactivator.

Wherein expand and/or microparticle rolls or slides on inclined surface between hardening period the inventive method initial, prevent such as generation during soft microparticle collides when being stirred to particulate damage.This meticulous processing has favourable effect for the global shape by the obtainable microparticle of the inventive method, and more specifically surface is favourable to this method for obtaining particulate integral spherical shape and/or smoothly (not having depression).Therefore the present invention allows to make the microparticle population, wherein less than 5%, more specifically less than 2% or have the most particularly even less than 1% population granule and to depart from (greater than 10%, more specifically greater than 5%) shape of integral spherical shape, and/or the injured surface that has depression and/or have other type.

Another aspect of the present invention relates to use microparticle of the present invention and is used for bioactivator is administered to mammal.

Microparticle of the present invention can be applied to various medical applications.Microparticle can be incorporated into and is used for parenteral and mucosal drug delivery in the drug ingredient, such as oral; Subcutaneous administration, intravenous administration, intra-arterial administration, intraperitoneal administration, muscle administration, vagina administration and rectally.

The superior properties of microparticle of the present invention makes that the size that they particularly are suitable for microparticle is crucial application, with the effect of avoiding not expecting.For for example thromboembolism therapy, for example in the treatment hysteromyoma, situation is such.Other application comprises the controlled release of bioactivator, because the fact of microparticle (by encapsulating more equably) size, this bioactivator is sent more accurately.

Thereby, the present invention relates to Therapeutic Method, this Therapeutic Method comprises that the microparticle of the present invention that will contain bioactivator (it is therapeutic compound) is administered to the patient who needs it.Additionally, the present invention relates to diagnostic method, this diagnostic method comprises that the microparticle of the present invention that will contain bioactivator (it is contrast agent or label) is administered to patient to be diagnosed.

Other layout that realizes system and method for the present invention is tangible to those skilled in the art.

To understand, although this paper at according to device description of the present invention preferred embodiment, particular configuration and configuration and material, can carry out various changes or adjustment in form and details and not deviate from scope and spirit of the present invention.

Example

Example 1: from 20 μ m nozzles injection PEGT/PBT copolymer emulsifying agent, frequency change.

1%PEG-PBT solution uses the microdroplet nozzle that immerses in the aqueous solution that comprises 0.3%pva 8/88 to come ink-jet.20 μ m nozzles are used.Ink-jet the pressure and 27.6,17 of 1.5bar, 12 and the frequency of 10kHz implemented 10 minutes.In aqueous solution, inclined surface is immersed and allows drop by rolling or slide through immersing apparent motion thus in another container.There, microparticle is waited until stable, supernatant be removed and the container water refills.Stir 1-2 hour with by be dissolved into contain remove dichloroethanes in the water after, on Ku Erte grain size analysis calculating instrument III, use 50 μ m apertures to measure microparticle.The mode diameter of volume weighted and number weight is 12.47,14.39,16.09 and 17.10 μ m.The volume weighted average diameter is 12.40,14.32,15.60 and 17.05 μ m.Distribution of sizes provides in Fig. 3.The width at peak shows that the granule (all countings of the counting of the Ku Erte between the 10-20 micron are taken into account) above 90% is in the meansigma methods of 0.5 μ m.

Example 2: the two emulsifying agents of injection PEGT/PBT copolymer.

1% water adds the solution of 1%PEG-PBT in dichloromethane to.Mixture uses Ultraturrax emulsifying 60 seconds, obtains the emulsifying agent of strong scattering.Use is immersed 30 μ m nozzles and in the frequency of 20.7kHz and the aux. pressure of 0.45bar this emulsifying agent ink-jet is comprised in the aqueous solution of 0.05%pva in first container.Described in example 1, drop is injected in the aqueous solution, is positioned at immersing on the inclined surface of first container.Drop rolls downwards gradually on the slope or slides in second container, waits until stable there.Supernatant be removed and the second container water refills.Stir 1-2 hour with by be dissolved into contain remove dichloromethane in the water after, on Ku Erte grain size analysis calculating instrument III, use 50 μ m apertures to measure microparticle.Use uniform pressure, frequency and reception fluid, PEG-PBT formulations prepared from solutions sample for reference from 1%.The distribution of sizes of two samples provides in Fig. 4 and the SEM photo of resulting granules provides in Fig. 5.For two emulsifying agents and for reference particles, the granule greater than 90% be number weighting diameter 4% in.

Example 3: two emulsifying agents of ink-jet PEG-PBT: contain fluorescin.

The preparation of example 2 is repeated, but combines fluorescin.Proteinic FITC modified mixture (the fluorescence molecule amount standard F3526 of Sigma-Aldrich) is used.1% the water that contains protein mixture adds the 1%PEG-PBT solution in the dichloromethane to.And then after the ink-jet, after following the tracks of particulate contraction and reaching final size, microscopically gathers photo, as shown in Figure 6.

Example 4: do not add the ink-jet of stabilizing agent.

In the dichloromethane 1%PEG-PBT solution use immerse 30 μ m nozzles at the aux. pressure of the frequency of 20.7kHz and 0.45bar and ink-jet in water, do not have any other additive.After cleaning, gather sample, obtain SEM photo shown in Figure 7.

Example 5: ink-jet plga and pla-peo.

The solution (A) that contains 0.35%pla-peo and 0.45%plga uses 20 microns nozzles, arrives in the aqueous 0.2%pva solution in frequency ink-jet in 10 minutes of 15kHz.After ink-jet, (left stirred) stirred to evaporate remaining solvent in twice of a product cleaning and a left side.Distribution of sizes is shown in Fig. 8.

Add 0.5% the water of the polypropylene glycol contain 3% surfactant and oxirane addition polymers (pluronic) F127 toward solution (A), and prepare emulsifying agent by using ultrasound.This obtains the emulsifying agent of water drop in polymer solution of strong scattering.Emulsifying agent use 20 microns nozzles the frequency of 15kHz in 10 minutes ink-jet in moisture 0.2%pva solution.After ink-jet, twice of a product cleaning and a left side are stirred to evaporate remaining solvent.Distribution of sizes is shown in Fig. 8.

Example 6: preparation contains the iodate liquid of capsule crop.

Suffering-2-the base 2,3 that contains 14 microns spheroids, the 5-Triiodobenzoic acid is by the solution in dichloroethanes prepares in 0.2%pva 8/88 solution with the speed ink-jet of 15kHz with plga and iodized oil.Particle size distribution is shown in Fig. 9, and microphotograph provides in Fig. 9 b.

Claims (8)

1. equipment that is used to produce polymer beads, described equipment comprises:

-the first reservoir (2) is used to keep emulsifying agent (3),

-the second reservoir (4) is used for keeping receiving fluid (5),

-jet module has at least one nozzle (6), and when this second reservoir (4) when being filled, this at least one nozzle (6) is immersed in this reception fluid (5), allow drop is directly injected in this reception fluid (5),

-inclined surface (7), be positioned in this second reservoir (4) or be associated with this second reservoir (4), make drop when in this reception fluid (5) that is ejected into from this nozzle (6) this second reservoir (4) along this inclined surface (7) motion, wherein this inclined surface (7) have vertically begin and with between 10 degree to gradual change slope that the angle between 45 degree finishes.

2. according to the equipment of claim 1, wherein this gradual change slope is to finish between 20 degree to the angle between 30 degree.

3. according to the equipment of claim 1, wherein this nozzle has the diameter less than 30 μ m.

4. method that is used for the single disperse water gelatin polymer microparticle of preparation size between 1 μ m and 100 μ m, wherein said microparticle comprises bioactivator, has wherein utilized the equipment according to claim 1, described method comprises the steps:

A) preparation comprises the emulsifying agent of the polymer that the hydrogel of described bioactivator forms,

B) use aux. pressure and applying frequency, flow the drop of the emulsifying agent of intravital this nozzle (6) ejection step (a) from being immersed in moisture reception, and,

C) by allowing this drop one time period of motion in this receives fluid, this time period guarantees that described drop removes and desolvates and water comes saturated, with this liquid drip expansion with harden into microparticle, wherein in step (c), this expansion and sclerosis are to carry out by allowing this granule to move downward from the inclined surface on slopes with at least 10 degree in this reception fluid.

5. according to the method for claim 4, wherein this polymer is an aquogel polymer.

6. according to the method for claim 4 or 5, wherein step (c) comprise allow this granule in this receptions fluid on described inclined surface one time period of motion, this time period guarantee to remove desolvate and water saturated.

7. the method any according to claim 4 to 6, wherein the length of this inclined surface and slope are arranged such that the time period of moving from the drop of this nozzle ejection along described inclined surface between 2 seconds to 60 seconds.

8. the method any according to claim 4 to 7, this method further are included in step (c) additional step (d) afterwards, wherein this granule further sclerosis by stirring.

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